[go: up one dir, main page]

WO2000031281A2 - Sequence d'adn codant un translocateur de glutamate/malate, plasmides, bacteries, levures et vegetaux contenant ledit transporteur - Google Patents

Sequence d'adn codant un translocateur de glutamate/malate, plasmides, bacteries, levures et vegetaux contenant ledit transporteur Download PDF

Info

Publication number
WO2000031281A2
WO2000031281A2 PCT/EP1999/008960 EP9908960W WO0031281A2 WO 2000031281 A2 WO2000031281 A2 WO 2000031281A2 EP 9908960 W EP9908960 W EP 9908960W WO 0031281 A2 WO0031281 A2 WO 0031281A2
Authority
WO
WIPO (PCT)
Prior art keywords
dna sequence
glutamate
malate
translocator
deletion
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/EP1999/008960
Other languages
German (de)
English (en)
Other versions
WO2000031281A3 (fr
Inventor
Ulf-Ingo FLÜGGE
Andreas Weber
Peter Westhoff
Uta Dressen
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
BASF SE
Original Assignee
BASF SE
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by BASF SE filed Critical BASF SE
Priority to EP99958097A priority Critical patent/EP1135510A2/fr
Priority to AU15554/00A priority patent/AU1555400A/en
Publication of WO2000031281A2 publication Critical patent/WO2000031281A2/fr
Publication of WO2000031281A3 publication Critical patent/WO2000031281A3/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
    • C12N15/8274—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for herbicide resistance
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8242—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
    • C12N15/8243—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine

Definitions

  • DNA - S FREQUENCY encoding a glutamate / malate translocator, plasmids, bacteria, yeasts and plants containing these transporters.
  • the present invention relates to DNA sequences from Sorghum bi color SEQ-ID No. 1, Flaveria bidentis SEQ-ID No. 3 and Spinacia oleracea SEQ-ID No. 5, which contain the coding region of a glutamate / malate translocator, the introduction of which into a plant genome changes the formation and transfer of carbon backbones for the fixation of nitrogen and the transfer of assimilated nitrogen in transgenic plants, plasmids, yeasts and bacteria containing this DNA Sequences, as well as transgenic plants, in which changes in the activity of the glutamate / malate translocator and thus changes in the nitrogen and carbon metabolism are caused by the introduction of the DNA sequences.
  • the invention further relates to transgenic plants which are affected by the change in the activity of the glutamate / malate translocator in their ability to photorespiration.
  • the invention also relates to the use of the described sequences of the glutamate / malate translocator for identifying related translocators from plants (angiosperms, gymsperms and algae) by hybridization with low stringency or by PCR techniques, and to the use of the glutamate / malate translocators as a target for herbicides.
  • the nitrogen supply to the plant must be influenced by fertilization.
  • the energy for nitrogen assimilation comes from the light reaction of photosynthesis or in roots and other non-green tissues from dissimilation and is not a limiting factor under normal field conditions.
  • 2-oxoglutarate (ketoglutarate) is the primary acceptor of reduced nitrogen in the glutamine synthetase / glutamine-2-oxoglutarate aminotransferase (GOGAT) reaction.
  • reduced nitrogen ammonium nitrogen
  • glutamate oxoglutarate aminotransferase (GOGAT; glutamate synthase) catalyzes the transfer of an amino acid using reduction equivalents.
  • the entire reaction sequence of the glutamine synthetase / GOGAT reaction is located in the stroma of the plastids of the plants. These organelles are enclosed by two lipid bilayer membranes, the outer molecular sieve character of which allows compounds up to a size of approx. 10 kDa to pass freely (Flügge and Benz,
  • the inner membrane is permeable to some smaller compounds such as water, carbon dioxide, oxygen and nitrite, but not to larger charged molecules such as glutamate or malate.
  • This key compound of the gl tamine synthetase / GOGAT reaction must be moved from the cytosol of the plant cell through a specific translocator across the inner plastid envelope membrane into the stroma of the plastid.
  • the transport of 2-oxoglutarate in the plastids takes place in exchange for malate from the
  • 2-oxoglutarate is imported into the chloroplast and the end product of the glutamine synthetase / GOGAT reaction, glutamate, is exported.
  • Glutamate serves as the preferred amino group donor in a whole series of transamination reactions, for example in the biosynthesis of the amino acids alanine or phenylalanine etc.
  • Most nitrogen-containing compounds in the plant, such as amino acids, nucleic acids or alkaloids, first require glutamate as the primary amino group donor in their biosynthetic pathway. Glutamate is also an important form of transport for organically bound nitrogen within the plant and is fed as such into the plant's leading tissue (the phloem).
  • the most common amino acids in phloem juice are usually glutamate, glutamine and aspartate.
  • Transporters are involved in the allocation of the photoassimilate (ie primarily sugar and amino acids) from the "source” - to the "sink” organs at a crucial point - such as the chloroplastid triose phosphate / phosphate translocator, the sucrose translocator, via which the sieve tubes loaded with the transport form of the photoassimilate sucrose and the glucose-6-phosphate / phosphate translocator of the amyloplasts. Changes in the activity of a specific membrane transporter can have a major impact on plant metabolic performance.
  • the glutamate / malate translocator of the plastid envelope membrane plays a key role in the intracellular distribution of the amino group acceptors and donors involved in nitrogen fixation. If it was possible to clone this translocator and to use the sequence obtained to modulate the activity of the translocator in plants, this opened the way to plants with a changed carbon / nitrogen ratio. This means that plants could be produced which, for example, contained fewer carbohydrates (sugar, starch, fats) and more organic nitrogen compounds (protein, amino acids, alkaloids). There is an urgent need for plants with such changed content substances, because the vast majority of people on earth depend on plant nutrition, with the consequence of a constant undersupply of proteins in these strata of the population.
  • plastid translocators require a corresponding presequence ("targeting" sequence) for correct addressing to the inner plastid membrane, which directs the attached mature protein to the plastids correctly (review article See: Keegstra et al., 1989, Annu. Rev. Plant Physiol. Plant Mol. Biol. 40: 471-501; Lubben et al., 1988, Photosynth. Res. 17: 173-194; diegge, 1990, J. Cell Sei. 96: 351-354).
  • the mature part (the part of the protein which remains after the presequence has been cleaved off by a specific protease) of the plastid envelope membrane proteins contains further information which is necessary for the specific insertion of the proteins into the membrane are responsible and prevent transport of the envelope membrane proteins over the envelope membrane into the plastid stroma or, in the case of the chloroplasts, the thylakoid membrane (Knight and Gray, 1995, Plant Cell 7: 1421-1432 or ' own investigations: Brink et al., 1995, J. Biol. Chem. 270: 20808-20815).
  • the protein / lipid interaction is important for correct insertion and the plastid envelope membrane differs fundamentally in its lipid composition from that of other cell organelles and also the bacteria (Joyard et al., 1991, Eur. J. Biochem. 199: 489-509), it is very unlikely that an insertion, albeit a small one, of a heterologous protein will then also be functional, that is to say that a transporter from other systems in the conformation and orientation corresponding to its function in the Plastid envelope membrane can be installed.
  • C 3 plants eg potato, tomato
  • C plants such as millet (Sorghum bicolor) or Flave ria bidentis
  • mesophyll and bundle sheath cells which are involved in the production of the photoassimilate are.
  • mesophyll cells the primary fixation of atmospheric carbon dioxide takes place with the formation of a C4 body (eg malate), which is then transported to the bundle sheath cells.
  • the corresponding cDNA insert was used as a probe for the subsequent screening of a cDNA library from Sorgum bicolor and a cDNA clone could be isolated which contains the complete cDNA (2088 base pairs) for the corresponding precursor protein with a molecular mass of 59 kDa contained.
  • Transformants expressing the corresponding plasmid were isolated. Transport experiments, carried out with membrane fractions of the yeast transformants reconstituted in artificial membranes, showed that this translocator catalyzes the transport of malate in exchange for glutamate (or aspartate). In contrast to the 2-oxoglutarate / malate translocator, this translocator also accepts amino acids (glutamate and aspartate) as counter-exchange substrates, see working examples 2 and 3. The transporter was given the name glutamate / malate translocator.
  • the present invention thus provides DNA sequences which originate from a plant genome and code for a plastid glutamate / malate translocator, the information contained in the nucleotide sequence leading to the formation and expression in plant cells of a ribonucleic acid and A glutamate / malate translocator activity can be introduced into the cells via this ribonucleic acid or an endogenous glutamate / malate translocator activity can be suppressed.
  • the invention particularly relates to the DNA sequences from sorghum bicolor (millet) with the nucleotide sequence SEQ-ID No. 1, a DNA sequence from Flaveria bidentis with the nucleotide sequence SEQ-ID No. 3 and a DNA sequence from Spinacia oleracea (spinach) with the nucleotide sequence SEQ-ID No. 5th
  • the invention further relates to DNA sequences which are identified by the sequence shown under SEQ ID No. 1, 3 or 5 mentioned DNA sequence or parts thereof or derivatives which are derived from these sequences by insertion, deletion or substitution, hybridize and code for a plastid protein which has the biological activity of a dicarboxylic acid translocator, in particular a glutamate / malate -Translocators owns.
  • the present invention relates to the use of the DNA sequence SEQ-ID No. 1, 3 or 5 or parts thereof or derivatives obtained by insertion, deletion or Substitution derived from these sequences for the transformation of pro- and eukaryotic cells.
  • the DNA sequence SEQ-ID No. 1, 3 or 5 are introduced into plasmids and are combined with control elements for expression in prokaryotic or eukaryotic cells, see exemplary embodiments 2 and 4.
  • control elements are, on the one hand, transcription promoters and, on the other hand, transcription terminators.
  • the plasmids can be used to transform eukaryotic cells with the aim of expressing a translatable messenger ribonucleic acid (RNA) which allows the synthesis of a plastid glutamate / malate translocator in the transformed cells, or with the aim of expressing a non-translatable, inversely oriented ("antisense"), messenger ribonucleic acid, which prevents the synthesis of the endogenous glutamate / malate translocators.
  • RNA messenger ribonucleic acid
  • antisense inversely oriented
  • RNA in accordance with the sequence of a plant glutamate / malate translocator according to the invention enables a change in plant nitrogen metabolism, the economic importance of which is that an improvement in the forwarding of glutamate from the cytosol of the plastids to a change in the ratio of Carbohydrates (sugar, starch, organic acids) and fats in favor of nitrogen compounds (amino acids, proteins, possibly alkaloids) takes place. Plants can thus be produced which are richer in valuable protein but have a lower content of carbohydrates and fats.
  • the transcriptional start area can be both native or homologous and foreign or heterologous to the host plant. Termination areas are interchangeable.
  • the DNA sequence of the transcription start and termination regions can be produced synthetically or obtained naturally or contain a mixture of synthetic and natural DNA components.
  • cloning vectors are available which contain a replication signal for E. coli and a marker which allows selection of the transformed cells. Examples of vectors are pBR322, pUC series, M13 mp series, pACYC 184 etc. Depending on the method of introducing desired genes into the plant, further DNA sequences may be required.
  • the Ti or Ri plasmid is used for the transformation of the plant, at least the right boundary, but often the right and left boundary of the Ti and Ri plasmid T-DNA must be added as a flank region to the genes to be introduced.
  • T-DNA for the transformation of plant cells has been intensively investigated and sufficiently described (Hoekema, in: The Binary Plant Vector System, Offset-drukkerij Kanters BV. Ablasserdam, 1985, Chapter V; Fraley et al., Critic. Rev. Plant Sc. 4: 1-46; An et al., 1985, EMBO J. 4: 277-287). Once the DNA has been integrated into the genome, it is generally stable there and is also retained in the offspring of the originally transformed cell.
  • Whole plants can then be regenerated in a suitable selection medium.
  • the plants obtained in this way can then be tested for the presence of the introduced DNA using conventional molecular biological methods.
  • These plants can be grown normally and crossed with plants that have the same transformed genetic makeup or other genetic makeup.
  • the resulting hybrid individuals have the corresponding phenotypic properties.
  • the DNA sequences SEQ-ID No. 1, 3 or 5 can also be introduced into plasmids and thereby provided with control elements for the expression in cells of fission yeasts, see embodiment example 2.
  • the introduction of the glutamate / malate translocator leads to a considerable one Increase in the activity of the glutamate / malate translocator measurable by reconstitution in artificial liposomes in the recombinant yeast cells. In recombinant yeast cells, a much higher malate transport activity can be demonstrated.
  • the DNA sequences SEQ-ID No. 1, 3 or 5 can also be introduced into plasmids which permit mutagenesis or a sequence change by recombination of DNA sequences in prokaryotic or eukaryotic systems. This allows the specificity of the glutamate / malate translocator to be changed e.g. towards an affinity for other dicarboxylic acids.
  • DNA sequence SEQ-ID No. 1, 3 or 5 (or parts or derivatives of this sequence) can be used according to standard methods (in particular hybridization screening of cDNA libraries with low stringency using the DNA sequence according to the invention or parts of the DNA sequences
  • the DNA sequence SEQ-ID No. 1, 3 or 5 contains areas in the translated protein that are able to specifically direct the protein synthesized in the cytoplasm on ribosomes to plastids and to prevent the occurrence of the protein in other membrane systems of the cell. This is in contrast to the known mitochondrial 2-oxoglutarate translocators, which do not contain such an addressing sequence.
  • the protein range which directs the protein encoded by the DNA sequence according to the invention to plastids lies within the first hundred amino acids of the protein, is not necessary for the transport function of the protein and is removed after the protein has been successfully inserted into the plastid envelope membrane.
  • the translocator protein By exchanging this "plastid targeting" sequence for one of the known “targeting" sequences, for example for mitochondria, the translocator protein could be eukaryotic in another membrane system. conduct cells and could possibly change the transport properties across the membrane.
  • the "plastid targeting" sequence of the glutamate / malate translocator or endogenous regions of the mature protein could be used to direct foreign proteins (eg bacterial transport proteins or transporters from yeast) into the plastids or into the plastid envelope membrane of plant cells.
  • the phage lambda ZAPII and the phagemid pBluescript II KS (pBSC) (Short et al., 1988, Nucl. Acids Res. 16: 7583- 7600) is used.
  • the vector SAP-E (Truenit et al., 1996, Plant Cell 8: 2169-2182) was used for the transformation of yeasts.
  • the E. coli strain DH5 ⁇ (Hanahan et al., 1983, J. Mol. Biol. 166: 557-580) was used for the pBluescriptKS (pBSC) phagemid and for SAP-E and pBinAR constructs.
  • the DNA was transferred into the agrobacteria by direct transformation using the method of Hofgen and Willmitzer (1988, Nucl. Acids Res. 16: 9877).
  • the plasmid DNA of transformed agrobacteria was isolated by the method of Birnboim and Doly (1979, Nucl. Acids Res. 7: 1513-1523) and, after a suitable restriction cleavage, analyzed for correctness and orientation by gel electrophoresis. 4.
  • DSM 12480 Phagemid pFbGMTl (GMTl sequence SEQ-ID No. 3 from Flaveria bidentis) (DSM 12478)
  • Poly (A + ) RNA was isolated from leaves of Sorghum bicolor, converted into double-stranded cDNA using a standard cDNA synthesis kit and cloned into the vector Lambda-ZAPII (Wyrich et al., 1998, Plant Mol. Biol. 37 : 319-335).
  • partial digestion with pectinases and cellulases from leaves of Sorghum bicolor prepared mesophyll and bundle sheath cells and from them poly (A + ) RNA. From the mesophyll or Bundle sheath RNA was produced with the help of reverse transcriptase and radioactive nucleotides, a hybridization probe with which the cDNA library was screened differentially.
  • the cDNA library was screened with the cDNA clone HHU51 as a hybridization probe. Lambda clones that reacted positively were cleaned using standard methods. The pBluescript phagemid with the cDNA insert coding for the glutamate / malate translocator was released by in vivo excision (Short & Sorge, 1992, Meth. Enzymol. 216: 495-208). The
  • the Sorghum bicolor (millet) cDNA was then used to generate the corresponding cDNA clones from Flaveria bidentis (clone pFbGMTl) (SEQ-ID No. 3) and Spinacia oleracea (spinach, clone pSoGMTl) (SEQ-ID No. 5) isolate.
  • SAP-E yeast Expression vector
  • the plasmid was transformed into S. pombe cells deficient in leucine synthesis deficient by LiCl / PEG (Ito et al., 1983, J. Bact. 153: 163-168). Transformants were selected by selection on minimal medium without leucine, since the SAP GMT construct gives the yeast cells the ability to grow on leucine-free medium.
  • Yeast cells transformed with the SAP-GMT plasmid were grown in minimal medium to an optical density at 600 nm of 1.0 and harvested by centrifugation at 3,000 x g for 5 min. The cells were then broken up by shaking vigorously with 1/2 vol (based on the cells) of glass beads and glass beads and cell fragments were separated off by centrifugation (600 g for 1 min). The supernatant was adjusted to a concentration of 0.5% (weight / volume) Triton X-100, the same volume of liposomes was added and the resulting proteoliposomes were immediately frozen in liquid nitrogen.
  • the liposomes were previously prepared by sonicating soybean phospholipid (20 mg / ml) for 10 min at 4 ° C in the presence of 200 mM tricine-NaOH (pH 7.6), 40 mM malate and 60 mM potassium gluconate. After thawing the proteoliposomes and re-sonicating the suspension with 10 pulses, the proteoliposomes were removed from the surrounding medium by size exclusion chromatography on Sephadex G-25, previously with 10 mM tricine-NaOH (pH 7.6), 100 mM sodium gluconate and 50 mM potassium gluconate had been equilibrated. The eluted proteoliposomes were used to measure malate transport activity. The measurement was carried out according to the "inhibitor-stop" method described by Menzlaff and wellgge (1993, Biochim. Biophys. Acta 1147: 13-18).
  • the malate transport activity in the SAP GMT transformants was compared to the malate transport activity of transformants that were only transformed with the vector SAP without the GMT insertion. It was shown that the malate transport activity in the SAP GMT transformants (measured in pmol transported 14 C-malate / mg protein per minute) was many times higher than in the SAP control transformants. It could also be shown that, in contrast to the already known 2-oxoglutarate / malate translocator, the glutamate / malate translocator is able to catalyze the transport of amino acids (glutamate / aspartate) in exchange for malate, see Table 1. Table 1
  • the recombinant glutamate / malate translocator and, for comparison, the recombinant 2-oxoglutarate / malate expressed in the split yeast Schizosaccharomyces pombe were reconstituted in liposomes which had been preloaded with the specified substrates.
  • the transport activities were determined as described by wellgge (Biochim. Biophys. Acta, 1992, 1110: 112-118) and are given as a percentage of the activity which was measured with malate-preloaded liposomes.
  • the 100% transport activity was 175 pmol / mg protein per minute.
  • pBSC-GMT phagemid pBluescript-GMT
  • the insert was isolated by restriction digestion with EcoRV and Smal and into the vector pBinAR
  • the transformants obtained were examined with the aid of Southern blot analyzes for the presence of the intact, non-rearranged chimeric gene. With the help of the "whole leaf reconstitution method" (Flügge and Weber, 1994, Planta, 194: 181-185) the malate transport activity in comparison to control transformants (transformed with vector pBinAR without insertion) was examined, as was the C / N- Ratio, photosynthesis rate, photorepiration and growth.

Landscapes

  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • Molecular Biology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • Plant Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Cell Biology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Botany (AREA)
  • Medicinal Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Nutrition Science (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)

Abstract

L'invention concerne des séquences d'ADN à base de sorghum bicolor (millet), flaveria bidentis et spinacia oleracea (épinard), contenant la région de codage d'un translocateur de glutamate/malate, dont l'introduction dans un génome végétal modifie la formation et la transmission de structures de base de carbone pour la fixation d'azote et la transmission de l'azote assimilé dans des plantes transgéniques. L'invention concerne en outre des plasmides, des levures et des bactéries contenant ces séquences d'ADN, ainsi que des plantes transgéniques dans lesquelles l'activité du translocateur de glutamate/malate, ainsi que des modifications du métabolisme de l'azote et du carbone sont induites par introduction de modifications des séquences d'ADN. De plus, l'invention concerne des plantes transgéniques influencées par la modification de l'activité du translocateur de glutamate/malate dans leur aptitude à la photorespiration. L'invention concerne également l'utilisation des séquences décrites du translocateur de glutamate/malate pour identifier des translocateur apparentés issus de végétaux (angiospermes, gymnospermes et algues), par hybridation à stringence réduite ou par des techniques de type PCR. L'invention concerne par ailleurs l'utilisation des translocateurs de glutamate/malate comme cibles («target») pour des herbicides.
PCT/EP1999/008960 1998-11-21 1999-11-22 Sequence d'adn codant un translocateur de glutamate/malate, plasmides, bacteries, levures et vegetaux contenant ledit transporteur Ceased WO2000031281A2 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP99958097A EP1135510A2 (fr) 1998-11-21 1999-11-22 Sequence d'adn codant un translocateur de glutamate/malate, plasmides, bacteries, levures et vegetaux contenant ledit transporteur
AU15554/00A AU1555400A (en) 1998-11-21 1999-11-22 Dna sequence encoding a glutamate/malate translocator, plasmids, bacteria, yeasts and plants containing said transporter

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE1998153778 DE19853778C1 (de) 1998-11-21 1998-11-21 DNA-Sequenzen kodierend einen Glutamat/Malat-Translokator, Plasmide Bakterien, Hefen und Pflanzen enthaltend diesen Transporter
DE19853778.6 1998-11-21

Publications (2)

Publication Number Publication Date
WO2000031281A2 true WO2000031281A2 (fr) 2000-06-02
WO2000031281A3 WO2000031281A3 (fr) 2000-07-27

Family

ID=7888573

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP1999/008960 Ceased WO2000031281A2 (fr) 1998-11-21 1999-11-22 Sequence d'adn codant un translocateur de glutamate/malate, plasmides, bacteries, levures et vegetaux contenant ledit transporteur

Country Status (4)

Country Link
EP (1) EP1135510A2 (fr)
AU (1) AU1555400A (fr)
DE (1) DE19853778C1 (fr)
WO (1) WO2000031281A2 (fr)

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002002798A3 (fr) * 2000-07-05 2003-04-17 Paradigm Genetics Inc Methodes et compositions permettant de moduler l'expression ou activite de la chorismate synthase et de la chorismate mutase dans les plantes
WO2003054207A3 (fr) * 2001-12-21 2004-01-29 Degussa Processus de fermentation pour la preparation d'acides amines l a l'aide de bacteries coryneformes
WO2006062971A3 (fr) * 2004-12-08 2006-11-30 Ceres Inc Modulation des teneurs en carbone dans les plantes
US7335760B2 (en) 2004-12-22 2008-02-26 Ceres, Inc. Nucleic acid sequences encoding zinc finger proteins
US7335510B2 (en) 2004-12-16 2008-02-26 Ceres, Inc. Modulating plant nitrogen levels
EP2275142A2 (fr) 2006-06-30 2011-01-19 Andre Koltermann Conjugués pour l'immunothérapie du cancer
US8222482B2 (en) 2006-01-26 2012-07-17 Ceres, Inc. Modulating plant oil levels
CN117535312A (zh) * 2024-01-10 2024-02-09 中国农业大学 一种液泡膜阴离子通道蛋白在调控玉米碳氮平衡中的应用

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE4420782C1 (de) * 1994-06-15 1995-08-17 Fluegge Ulf Ingo Prof Dr DNA-Sequenz, kodierend einen 2-Oxoglutarat/Malat-Translokator, Plasmide, Bakterien, Hefen und Pflanzen enthaltend diesen Transporter

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002002798A3 (fr) * 2000-07-05 2003-04-17 Paradigm Genetics Inc Methodes et compositions permettant de moduler l'expression ou activite de la chorismate synthase et de la chorismate mutase dans les plantes
US6800459B2 (en) 2000-07-05 2004-10-05 Paradigm Genetics, Inc. Methods and compositions for the modulation of chorismate synthase and chorismate mutase expression or activity in plants
WO2003054207A3 (fr) * 2001-12-21 2004-01-29 Degussa Processus de fermentation pour la preparation d'acides amines l a l'aide de bacteries coryneformes
WO2006062971A3 (fr) * 2004-12-08 2006-11-30 Ceres Inc Modulation des teneurs en carbone dans les plantes
US7329797B2 (en) 2004-12-08 2008-02-12 Ceres, Inc. Modulating plant carbon levels
US8299320B2 (en) 2004-12-08 2012-10-30 Ceres, Inc. Modulating plant carbon levels
US7335510B2 (en) 2004-12-16 2008-02-26 Ceres, Inc. Modulating plant nitrogen levels
US7335760B2 (en) 2004-12-22 2008-02-26 Ceres, Inc. Nucleic acid sequences encoding zinc finger proteins
US8222482B2 (en) 2006-01-26 2012-07-17 Ceres, Inc. Modulating plant oil levels
EP2275142A2 (fr) 2006-06-30 2011-01-19 Andre Koltermann Conjugués pour l'immunothérapie du cancer
CN117535312A (zh) * 2024-01-10 2024-02-09 中国农业大学 一种液泡膜阴离子通道蛋白在调控玉米碳氮平衡中的应用
CN117535312B (zh) * 2024-01-10 2024-04-09 中国农业大学 一种液泡膜阴离子通道蛋白在调控玉米碳氮平衡中的应用

Also Published As

Publication number Publication date
EP1135510A2 (fr) 2001-09-26
AU1555400A (en) 2000-06-13
WO2000031281A3 (fr) 2000-07-27
DE19853778C1 (de) 2000-09-21

Similar Documents

Publication Publication Date Title
EP0765393B1 (fr) Molecules d'adn codant pour un translocateur plastidial de 2-oxoglutarate/malate
EP0375092A1 (fr) Régulation de transcription spécifique pour des tubercules de pomme de terre
DE69333180T2 (de) Für oligosaccharidetransporter kodierende dna sequenzen
DE4004800A1 (de) Im habitus und ertrag veraenderte transgene pflanzen
EP3129394B1 (fr) Protéines tonoplastiques antiporteurs sucre-protons, et leur utilisations pour augmenter la concentration de saccharose d'un organe de stockage de saccharose chez les végétaux
DE4207358A1 (de) Expressionskassette und plasmide zur schliesszellenspezifischen expression und ihre verwendung zur herstellung transgener pflanzenzellen und pflanzen
DE4337597C2 (de) DNA-Sequenzen für Ammoniumtransporter, Plasmide, Bakterien, Hefen, Pflanzenzellen und Pflanzen enthaltend den Transporter
DE69326770T2 (de) Für einen amnosäauretransporter kodierende dna-sequenzen, plasmide, bakterien,hefen, und pflanzen die einer transporter enthalten sowie ihre verwendung
DE4444460A1 (de) Verfahren zur Steigerung des Ertrags sowie zur Veränderung des Blühverhaltens bei Pflanzen
DE19853778C1 (de) DNA-Sequenzen kodierend einen Glutamat/Malat-Translokator, Plasmide Bakterien, Hefen und Pflanzen enthaltend diesen Transporter
DE69325264T2 (de) Methode zur erhöhung der frühreife bei pflanzen und/oder reduzierung des nitratgehaltes einer pflanze
DE19734218A1 (de) Verfahren zur Ertragssteigerung in Pflanzen
DE19732926C2 (de) DNA-Sequenzen, kodierend einen Glukose-6-phosphat-Phosphat-Translokator, sowie Plasmide, Bakterien, Hefen und Pflanzen, enthaltend diesen Transporter
DE19600357C1 (de) DNA-Sequenz kodierend einen Phosphoenolpyruvat-Phosphat-Translokator, Plasmide, Bakterien, Hefen und Pflanzen enthaltend diesen Transporter
EP1276882A2 (fr) Procede de modification genetique d'une plante
EP1307561B1 (fr) Acides nucleiques a l'aide desquels peuvent etre produits des vegetaux a teneur en metabolites modifiee
EP1346054B1 (fr) Acides nucleiques codant pour des invertases vacuolaires, cellules vegetales et plantes contenant ces acides nucleiques, ainsi que leur utilisation
DE10313795A1 (de) Veränderte PPase-Expression in Zuckerrübe
EP1070120A1 (fr) Adenylique-desaminase
DE19914792A1 (de) Metabolische Selektionsmarker für Pflanzen
DE19826444C1 (de) DNA-Sequenzen kodierend einen Glukose-Translokator, Plasmide, Bakterien, Hefen und Pflanzen enthaltend diesen Transporter
DE10050233A1 (de) Verfahren zur genetischen Modifizierung einer Pflanze
DE4408629A1 (de) Verfahren zur Inhibierung der Blütenbildung in Pflanzen
DE4438821A1 (de) Verfahren zur Inhibierung sowie zur Induktion der Blütenbildung in Pflanzen (III)
DE4435366A1 (de) Verfahren zur Inhibierung der Blütenbildung in Pflanzen

Legal Events

Date Code Title Description
ENP Entry into the national phase

Ref country code: AU

Ref document number: 2000 15554

Kind code of ref document: A

Format of ref document f/p: F

AK Designated states

Kind code of ref document: A2

Designated state(s): AU CA IL JP RU US

AL Designated countries for regional patents

Kind code of ref document: A2

Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE

121 Ep: the epo has been informed by wipo that ep was designated in this application
AK Designated states

Kind code of ref document: A3

Designated state(s): AU CA IL JP RU US

AL Designated countries for regional patents

Kind code of ref document: A3

Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
WWE Wipo information: entry into national phase

Ref document number: 1999958097

Country of ref document: EP

WWP Wipo information: published in national office

Ref document number: 1999958097

Country of ref document: EP

WWW Wipo information: withdrawn in national office

Ref document number: 1999958097

Country of ref document: EP